蛋白质、核酸标记和检测
 
DIG标记及检测试剂盒
 
DIG(地高辛)DNA标记及检测试剂盒
 
DIG DNA labeling and Detection Kit：Random primed DNA labeling with digoxigenin-dUTP, alkali-labile and detection of hybrids by enzyme immunoassay
 
原理：用DIG-dUTP为底物，随机掺入法进行DNA标记，通过酶联免疫杂交的方法检测。该试剂盒可以标记25次反应，每次标记的DNA量为10ug-20ug，并可做50张10×10cm膜的杂交和检测。

应用：非放射性DNA标记及检测试剂盒以检测0.1pg的同源DNA，能检测1ug哺乳动物DNA中的单拷贝基因。跟放射性标记系统比较，此试剂盒能快速得到实验结果，而且消除了放射性同位素的不安全问题。从DNA标记和杂交能见到检测结果，整个实验过程24小时内能完成。这试剂盒的灵敏度与特异性使它适用于所有的杂交技术以替代放射性同位素标记和放射自显影术。这个标记方法和检测系统适用于DNA印迹转移、菌落杂交、噬斑杂交及原位杂交。
 
产品号
 产品规格
 厂商
 价格
 
1093657

11093657910
 Kit：1 kit (25 labeling reactions and detection of 50 blots)
 Roche
 7447
 


DIG DNA Labeling and Detection Kit

Application

Random-primed labeling of DNA probes with DIG-11-dUTP, alkali-labile and color detection of DIG-labeled hybrids. DIG-labeled probes are used for hybridization in Southern, northern, and dot blots, as well as for colony and plaque screening.
DIG-labeled hybrids are detected with an anti-DIG-alkaline phosphatase conjugate and the substrates NBT (nitroblue tetrazolium salt) and BCIP (5-bromo-4-chloro-3-indolyl phosphate, toluidinium salt), which give a light-blue precipitate.

Product Description

Sensitivity and specificity: The standard labeling reaction with 1 g template will give a yield of 260 ng DIG-labeled DNA after a 1-hour incubation at 37°C, or 780 ng after a 20-hour incubation. The sensitivity of DIG detection depends on the concentration of the DIG-labeled probe in the hybridization reaction, and the duration of the color reaction.


Table 1: Sensitivity of DIG detection.

Contents

1.       Unlabeled Control DNA 1, 20 l, 100 g/ml 

2.       Unlabeled Control DNA 2, 20 l, 200 g/ml 

3.       DNA Dilution Buffer, 2 x 1 ml 

4.       DIG-labeled Control DNA, 50 l, 5.2 g/ml 

5.       10x Hexanucleotide Mix, 50 l 

6.       10x dTP Labeling Mixture, 50 l 

7.       Klenow Enzyme, labeling grade, 25 l, 2 U/l 

8.       Anti-digoxigenin-AP-conjugate, 200 l, 750 U/ml 

9.       NBT/BCIP Concentrated Stock Solution, 10 x 1 ml 

10.    Blocking Reagent, 2 x 50 g